[Rapid diagnosis of paroxysmal nocturnal hemoglobinuria by gel test agglutination]. / Diagnostic rapide des hémoglobinuries nocturnes paroxystiques par agglutination en gel.

Murine monoclonal antibodies (MoAbs) directed against DAF (Decay Accelerating Factor, CD55 antigen) and MIRL (Membrane Inhibitor of Reactive Lysis, CD59 antigen) were used to identify the affected red cells (CD55-/CD59-) of PNH patients. MoAbs NaM16-4D3 (CD55, IgG2a) and NaM77-1E5 (CD59, IgG3) weakly agglutinate red cells and represent powerful tools to quantitate normal (PNHI) and abnormal (PNHII and PNHIII) cells from PNH patients by indirect flow cytometry. MoAbs NaM125-7H10 (CD55) and NaM123-6G12 (CD59), both IgM, were selected for their agglutinating properties and used for the separation of PNHI from PNHII and PNHIII red cells by the gel test technology. From analysis of artificial mixtures of DAF+ and DAF- cells, a direct relationship was established between fluorescent cells detected by flow cytometry, and erythrocytes agglutinated in microtyping cards. The method was further confirmed by analysis of ten blood samples from PHN patients and represent an alternative to classical hemolysis tests. On the basis of our experience we propose the following for the diagnosis of PNH: 1) agglutination test with NaCl microtyping cards using IgM CD55 and CD59; 2) flow cytometry analysis for accurate quantitation of CD55-/CD59- red cells.

[Paroxysmal nocturnal hemoglobinuria. Diagnosis aided by a monoclonal antibody directed against the decay accelerating factor glycoprotein]. / Hémoglobinurie nocturne paroxystique. Diagnostic à l'aide d'un anticorps monoclonal dirigé contre la glycoprotéine decay accelerating factor.

The laboratory diagnosis of paroxysmal nocturnal haemoglobinuria (also called Marchiafava-Micheli disease) is based on the sensitivity of the patient's red cells to complement-induced lysis. In view of the clonal expression of the disease, haemolysis tests are difficult to interpret when the abnormal red cell population is small. The sensitivity of abnormal red cells to haemolysis is due to the absence of proteins attached to the cell membrane by a phosphatidyl-inositol link, which intervene in the regulation of the complement-induced lysis mechanism. Using a monoclonal antibody directed against one of these proteins, the decay accelerating factor (DAF, protein CD 55), makes it possible to diagnose paroxysmal nocturnal haemoglobinuria. DAF expression on patients' blood cells was measured by quantitative agglutination and by indirect flow cytometry. The agglutination test using polybren is a fast detection method, but it may be uninterpretable, notably in cases with positive antiglobulin (Coombs') test. In contrast, DAF expression measured by indirect flow cytometry correlates perfectly with measurement of red cell sensitivity by haemolysis tests. Using the monoclonal antibody by indirect flow cytometry is the method of choice to confirm the diagnosis of paroxysmal nocturnal haemoglobinuria and to measure the proportions of normal and abnormal red cells in case of haematological disorder.

[Characterization of murine monoclonal antibodies against fetal erythrocytes]. / Caractérisation d'anticorps monoclonaux murins dirigés contre les érythrocytes foetaux.

Balb/c mice were immunized against papain-treated fetal erythrocytes and splenocytes were fused with Sp2/0-Ag-14 myeloma cells. Several hybrids secreting antibodies directed against antigenic determinants predominantly exposed on fetal and cord cells were selected and cloned twice. Antibodies NaM61-1A2 and NaM61-768 (IgM class) were shown to be specific for an endo-beta-galactosidase-sensitive oligosaccharide chain. The antigen, strongly expressed on fetal and cord cells, was identified as the i blood group antigen. The antibodies represent powerful blood group reagents to be use in conventional agglutination techniques as well as in the gel typing system and in indirect flow cytometry. The antibody NaM46-4A8 (IgG class) is specific for an antigenic structure expressed on fetal cells and accessible only after papain, ficin, bromelin and endo-beta galactosidase treatment. The antigen was not identified.

[Clinical hemorheologic study of 65 inflammatory syndromes explored using standard biologic examination and total blood filtration test]. / Etude hémorhéologique clinique de 65 syndromes inflammatoires explorés par bilan biologique standard et test de filtration de sang total.

The hemorrheologic impact of an inflammatory syndrome was investigated in a prospective biological study of 65 patients by using an erythrocyte filtration test on total blood. Impact was assessed by a statistical study of correlations between relative filtration time (RFT), inflammation-reactive proteins (IRP) and the different parameters of the hemogram. A stepwise multiple regression test (SMRT) indicated the dominant role of fibrinogen in determining acceleration of the sedimentation rate (p less than 10(-3) and lengthening of RFT (p less than 10(-3). Moreover, RFT appeared to be very significantly correlated with polymorphonuclears (R = 0.55; p less than 10(-3] hematocrit as measured by the centrifugation technique (R = 0.34; p = 0.005) and corpuscular concentration in hemoglobin (CCHM) (R = 0.30; p = 0.01). A chart predictive of microcirculatory risk was deduced from the study based on fibrinogen values and hematocrit. However, the relatively low rate of reduction in the total variance observed during SMRT suggests the quite relative role of the studied parameters in erythrocyte filtration disturbances during an inflammatory syndrome.

[Migration of polynuclear neutrophils in vivo. Standardization of the technic]. / Contribution à l'étude de la migration des polynucléaires neutrophiles in vivo. Standardisation de la technique.

The migration of neutrophils across a skin barrier is a commonly employed method for the study of the inflammatory response. This report aims at comparing the leukocyte migration in the presence of two different chemoattractants : autologous serum and pool of serum. The reproducibility of this method has proved to be increasing by using duplicate skin chambers in 30 healty volunteers. The neutrophil migration with the pool of serum appears more homogeneous than with autologous serum. These results permit to obtain a standard curve of the neutrophil migration using a pool of serum.

[Antibody-dependent cell-mediated cytotoxicity: a method of predicting the severity of fetomaternal alloimmunization in the Rhesus system? A retrospective study of 32 cases]. / La cytotoxicité anticorps dépendante à médiation cellulaire (A.D.C.C.): un moyen de prévoir la gravité des allo-immunisations foeto-maternelles dans le système Rhésus? Etude rétrospective à partir de 32 cas.

This retrospective study has as its object to find out whether A.D.C.C. has advantages over other techniques that are usually used, namely titres or levels of antibodies. There is no correlation between A.D.C.C. and the levels or titres of antibodies in the 32 cases that were studied. When the level of bilirubin at birth is taken as the criterion of the severity of the illness, A.D.C.C. is the only method that gives a significant correlation with the degree of severity of the condition of the infant (p = 0.01). If the need for treatment is taken as a criterion of the severity of the condition (ultra-violet light or replacement-transfusion), A.D.C.C. shows up as being the better technique. But if one only looks at very anaemic infants the significance becomes much greater (p = 0.001).

Plasma exchange in early kidney graft rejection associated with anti-donor antibodies.

Patients with early rejection of kidney allografts associated with anti-donor antibodies have been randomized in two groups which received, respectively, either the conventional corticosteroid/azathioprine treatment or extensive plasma exchanges (PE) plus the conventional treatment. Data on the monitoring of anti-T or anti-B donor lymphocytes, as well as anti-panel or autoreactive cytotoxicity are described. Although the titer of anti-donor antibodies is decreased in the PE-treated group there is no sustained improvement of graft function compared to the control group. Thus, in these stereotyped rejection episodes, which are likely to be antibody mediated, there is no significant effect of extensive and early plasma exchange.

Systematic transfusion in hemodialyzed patients awaiting grafts: kinetics of anti-t and b lymphocyte immunization and its incidence on graft function.

Since June 1977, a systematic blood transfusion (BT) policy (160 ml of leukocyte-poor washed erythrocytes given every 6 months) has been applied to 126 hemodialyzed patients awaiting a first kidney graft. Only patients who had anti-T or B lymphocyte (T or BLY) antibodies (Ab) killing fewer than 10 or 20% of the panel cells, respectively, entered the protocol. Screening of anti-T and BLY was performed 8, 15, and 21 days after each BT. Patients were removed from the protocol if they developed Ab against more than 10 or 20% of the T or BLY panel cells. The cumulative immunization (all Ab types) averaged 90% after four BTs. Anti-BLY (63%) were more frequent than anti-TLY (49%) after for BTs. No anti-HLA-DR specificity could be attributed to the anti-BLY, whereas 20% of the anti-TLY displayed a particular anti-HLA-A,B specificity. Patients that had had BTs or pregnancies before entering the protocol had a higher degree of immunization. The kinetics of the anti-B or TLY pattern differed greatly both at the level of their detection after 8, 15, and 21 days following one BT and in their development after repeated BTs. Forty-three patients received transplants at various stages of the protocol. Recipients grafted without Ab had the best graft outcome (87 versus 66 actuarial percentage at 3 months), even though their HLA (A,B and DR) matching was inferior. There was no significant difference in recipients who had different subgroups of Ab. These data indicate that immunization is very high even after a few BTs when careful controls are performed and they suggest that BTs do not act via active enhancement.

HLA antigens and Berger's disease.

We have studied the frequencies of HLA-A, -B antigens in 73 Berger's disease patients, plus HLA-DR antigens in 35 of them, and compared the percentages of antigens frequencies with those of a local and national panel. This study does not confirm the positive associations with HLA-Bw35 or HLA-B12 which have been previously reported. The HLA-DR typing only showed increased frequency of blanks in the patients (P smaller than 0.01, but no significant corr.P). Patients with Berger's disease and renal failure have a higher (but still not significant) HLA-Bw35 frequency than those without renal failure. The reasons for the discrepancy between our group and others are analysed.

[Blood filtrability after serious burn trauma (author's transl)]. / La filtration du sang chez les brûlés graves.

We previously stated that blood filtrability was very decreased during 30 to 60 days after burn traums. After one week there is a significant relationship between the filtrability of the RBC and the burnt area. In the beginning of the phenomenon many factors seem to be involved: --intra-erythrocyte ATP variations,--hemodynamic disorders of the early shock,--disturbances of enzymatic systems of shock secondary-effect,--reactions related to the protidic catabolism. The elucidation of this mechanism though not essentially univocal, may be important of state for the therapeutic protocol in terms of tissus oxygenation, healing and grafting.

[Coagulometer H21: an automatic apparatus for coagulation study (author's transl)]. / Le coagulomètre H21: appareil automatique pour l'étude de la coagulation.

We describe and present a new automatic device to measure the clotting time of a plasma plus reagent mixture by recording the time of transformation : solution to gel. The apparatus is able to perform 50 tests without manual help. Indeed it automatically carries out:--the dispatching of the tested plasma,--the dispatching of the various reagents,--the recording of the clotting time of the sample,--the processing of the data through greater a printer. No contamination is allowed to occur since the tubing and the plates containing the sample are disposable. We obtain accuracy than with the manual method with a coefficient of variation below 3%.

[Groupamatic 360 C1 and transfusion safety]. / Groupamatic 360 C1 et recherche de la sécurité transfusionnelle.

Presentation of an evaluation program of patient blood grouping through Groupamatic, with final report printed out under the control of the management computer of the blood center. The flow chart is composed of three main steps:--information tracking on a two-part card,--duplicated determination of blood groups and phenotypes by Groupamatic,--correlation check between the two runs by the management computer, followed by the print out of the blood grouping results. Print out always includes two STEPS:--for the first blood grouping: self adhesive label on a color form and on the transfusion record,--for the second blood grouping: definite blood group card and check label for transfusion record,--for antibody screening and pretransfusion check up, results and label with a summary taped on. Transfusion file should always follow the patient and allows to write his transfusion "past" just by sticking on the I.D. numbers of transfused products. Furthermore, the management computer prints out:--the result log book.--a daily updated alphanumeric listing.

[Groupamatic 360 C1 and automated blood donor processing in a transfusion center]. / Groupamatic 360 C1 et gestion automatisée du circuit des donneurs dans un centre de transfusion.

Automation of donor management flow path is controlled by: --a 3 slip "port a punch" card, --the groupamatic unit with a result sorted out on punch paper tape, --the management computer off line connected to groupamatic. Data tracking at blood collection time is made by punching a card with the donor card used as a master card. Groupamatic performs: --a standard blood grouping with one run for registered donors and two runs for new donors, --a phenotyping with two runs, --a screening of irregular antibodies. Themanagement computer checks the correlation between the data of the two runs or the data of a single run and that of previous file. It updates the data resident in the central file and prints out: --the controls of the different blood group for the red cell panel, --The listing of error messages, --The listing of emergency call up, --The listing of collected blood units when arrived at the blood center, with quantitative and qualitative information such as: number of blood, units collected, donor addresses, etc., --Statistics, --Donor cards, --Diplomas.

Genetic variants of serum alpha-1-antitrypsin (Pi types) in Bretons.

The results of Pi typing on 280 Bretons from Morbihan (Southern Brittany) are reported. 6 phenotypes and 5 alleles have been found in this study. Pi M is the most frequent as in other populations. Pi S and Pi F apears as the two main variants in population genetics.

Use of modified automated polybrene test for screening IgG sensitized RBC in routine laboratory.

An automatic method for the screening of RBC sensitized by IgG, using the polybrene reagent was utilized. We compared data drawn from the analysis of the automatic routine test in 4799 patients with the results obtained by the direct anti-IgG antiglobulin test. We give evidence for the highest sensitivity of the polybrene test with a good overlap of the results of two methods.

[The beneficial role of blood transfusion and preganancy in the survival of renal allografts]. / Rôle bénéfique des transfusions sanguines et des grossesses dans la survie des allogreffes rénales.

This analysis involves 93 cases of transplants of non-related cadaver kidneys. The result was better (p = 0.02 at 1 year) in recipients transfused before the graft (73 cases) than in those who had not been trasfused (20 cases). However, this latter group had a shorter average period of haemodialysis (p = 0.05 than the first. The benefits of pre-immunisation appear from 1 to 2 transfusions or 1 pregnancy onwards (p = 0.02 at 1 year). Subjects transfused during the 6 months prior to the transplant had a graft which was functional more often than those who had been more than 6 months before the operation (p = 0.01 at 6 months). These results would be in favour of routine small quantity transfusions, approximately every six months in patients undergoing haemodialysis and on a waiting list for transplantation.